Aims: Clinically assess post-operative pain following root canal preparation using various single-file systems. Material and Methods: For this study, thirty patients between the ages of 20 and 40 who needed a routine root canal treatment for a tooth with a single root canal were chosen. Cases were randomly divided into three equal groups (n = 10). Group 1: Hyflex EDM; Group 2: XP-endo Shaper; and Group 3: Primary Wave One Gold. Root canal treatment was done, and the postoperative pain severity was evaluated using a Visual Analogue Scale (VAS) at 24, 72 hours, and 7 days after treatment. Results: When compared to the Hyflex EDM and Wave One Gold groups, the incidence of postoperative pain was greater in the XPendo Shaper group, but without statistically significant differences. In all groups, the incidence of reported postoperative pain decreased with time. Conclusions: The use of different tested root canal instruments has no significant impact on postoperative pain. All groups experienced a progressive decrease in postoperative pain intensity.